This study aimed to determine whether actinomycetes could contribute to the odours occurring in Lake Lotus, China. In total, 55 actinomycete strains were isolated from sediments in Lake Lotus and their odorous metabolites, geosmin and 2-methylisoborneol (2-MIB), were identified using HSPME-GC-MS. Results showed that 24, 23 and 34 strains produced geosmin and/or 2-MIB in Gause, TSB and M liquid media, respectively. Of odour-producing actinomycetes, most could produce geosmin and some produced both metabolites, while few of them produced only 2-MIB. Six isolates with high-level odour were selected for further investigation. Their biomass and odour-producing abilities were monitored in both the slants and liquid media. The results suggest that TSB was the most suitable medium for the growth of mycelium; Gause and M slant supported good production of spores, while M liquid medium was the most favourable for the production of both geosmin and 2-MIB. Those strains that produced geosmin only were less influenced than those that produced both geosmin and 2-MIB under shaking conditions. The results indicate that actinomycetes from sediments should be taken into consideration when off-flavours occur in water columns. According to the 16S rRNA sequences, six actinomycetes were classified in the Streptomyces.